Reinterpretation of tuberculate cervical vertebrae of Eocene birds as an exceptional anti-predator adaptation against the mammalian craniocervical killing bite.

We report avian cervical vertebrae from the Quercy fissure fillings in France, which are densely covered with villi-like tubercles. Two of these vertebrae stem from a late Eocene site, another lacks exact stratigraphic data. Similar cervical vertebrae occur in avian species from Eocene fossils sites in Germany and the United Kingdom, but the new fossils are the only three-dimensionally preserved vertebrae with pronounced surface sculpturing. So far, the evolutionary significance of this highly bizarre morphology, which is unknown from extant birds, remained elusive, and even a pathological origin was considered. We note the occurrence of similar structures on the skull of the extant African rodent Lophiomys and detail that the tubercles represent true osteological features and characterize a distinctive clade of Eocene birds (Perplexicervicidae). Micro-computed tomography (µCT) shows the tubercles to be associated with osteosclerosis of the cervical vertebrae, which have a very thick cortex and much fewer trabecles and pneumatic spaces than the cervicals of most extant birds aside from some specialized divers. This unusual morphology is likely to have served for strengthening the vertebral spine in the neck region, and we hypothesize that it represents an anti-predator adaptation against the craniocervical killing bite ("neck bite") that evolved in some groups of mammalian predators. Tuberculate vertebrae are only known from the Eocene of Central Europe, which featured a low predation pressure on birds during that geological epoch, as is evidenced by high numbers of flightless avian species. Strengthening of the cranialmost neck vertebrae would have mitigated attacks by smaller predators with weak bite forces, and we interpret these vertebral specializations as the first evidence of "internal bony armor" in birds.

Frontoparietal Bone in Extinct Palaeobatrachidae (Anura): Its Variation and Taxonomic Value.

Palaeobatrachidae are extinct frogs from Europe closely related to the Gondwanan Pipidae, which includes Xenopus. Their frontoparietal is a distinctive skeletal element which has served as a basis for establishing the genus Albionbatrachus. Because little was known about developmental and individual variation of the frontoparietal, and its usefulness in delimiting genera and species has sometimes been doubted, we investigate its structure in Palaeobatrachus and Albionbatrachus by means of X-ray high resolution computer tomography (micro-CT). To infer the scope of variation present in the fossil specimens, we also examined developmental and interspecific variation in extant Xenopus. In adults of extinct taxa, the internal structure of the frontoparietal bone consists of a superficial and a basal layer of compact bone, with a middle layer of cancellous bone between them, much as in early amphibians. In Albionbatrachus, the layer of cancellous bone, consisting of small and large cavities, was connected with the dorsal, sculptured surface of the bone by a system of narrow canals; in Palaeobatrachus, the layer of cancellous bone and the canals connecting this layer with the dorsal surface of the frontoparietal were reduced. The situation in Palaeobatrachus robustus from the lower Miocene of France is intermediate-while external features support assignment to Palaeobatrachus, the inner structure is similar to that in Albionbatrachus. It may be hypothesized that sculptured frontoparietals with a well-developed layer of cancellous (i.e., vascularized) bone may indicate adaptation to a more terrestrial way of life, whereas a reduced cancellous layer might indicate a permanent water dweller.

Do all frogs swim alike? The effect of ecological specialization on swimming kinematics in frogs.

Frog locomotion has attracted wide scientific interest because of the unusual and derived morphology of the frog pelvic girdle and hind limb. Previous authors have suggested that the design of the frog locomotor system evolved towards a specialized jumping morphology early in the radiation of the group. However, data on locomotion in frogs are biased towards a few groups and most of the ecological and functional diversity remains unexplored. Here, we examine the kinematics of swimming in eight species of frog with different ecologies. We use cineradiography to quantify movements of skeletal elements from the entire appendicular skeleton. Our results show that species with different ecologies do differ in the kinematics of swimming, with the speed of limb extension and especially the kinematics of the midfoot being different. Our results moreover suggest that this is not a phylogenetic effect because species from different clades with similar ecologies converge on the same swimming kinematics. We conclude that it is important to analyze frog locomotion in a broader ecological and evolutionary context if one is to understand the evolutionary origins of this behavior.

Anurans from the Lower Cretaceous Jehol Group of western Liaoning, China.

BACKGROUND: To date, the Lower Cretaceous Jehol Group of western Liaoning, China has yielded five monotypic genera of anurans, including Liaobatrachus grabaui, Callobatrachus sanyanensis, Mesophryne beipiaoensis, Dalianbatrachus mengi, and Yizhoubatrachus macilentus. However, the validity and distinctness of these taxa have been questioned. METHODOLOGY/PRINCIPAL FINDING: We provide a comprehensive analysis of the Jehol frogs that includes a re-examination of the published taxa as well as an examination of a number of new specimens that have been collected over the past 10 years. The results show that the five previously named taxa can be referred to three species of one genus-Liaobatrachus grabaui, L. beipiaoensis comb. nov. and L. macilentus comb. nov.. The diagnosis of Liaobatrachus is revised, and a new diagnosis is provided for each species of this genus. We also establish Liaobatrachus zhaoi sp. nov., on the basis of a dozen well-preserved specimens from a new locality. This taxon is distinguished by a unique combination of characteristics, including relatively long hind limbs, a rounded rather than triangular acetabulum, and a gradually-tapering cultriform process of the parasphenoid. In addition, an unnamed frog from a higher horizon, which has narrow sacral diapophyses and particularly long legs, is different from Liaobatrachus and represents another form of anuran in the Jehol Biota. CONCLUSION/SIGNIFICANCE: Comparisons with other Mesozoic and extant anurans and the primary phylogenetic analysis both suggest that Liaobatrachus is a member of the anuran crown-group and forms a polytomy with leiopelmatids (Ascaphus and Leiopelma) and the remaining crown-group anurans (Lalagobatrachia).

Development of the ethmoidal structures of the endocranium in Discoglossus pictus (Anura: Discoglossidae).

We use histological techniques and computer-aided three-dimensional reconstructions made from serial histological sections to describe the ontogeny of the ethmoidal endocranium of discoglossid frog Discoglossus pictus. We identify a pattern of development for the suprarostral cartilage that differs from previous findings and probably represents the ancestral anuran pattern. The nasal cartilages, including the inferior prenasal cartilage, are de novo adult structures. The only larva-derived structures of the adult nasal capsules are the posterior aspects of the solum nasi and septum nasi. We also identify patterns of development for the ethmoid plate and postnasal wall that occur during early in ontogenesis. These patterns are associated with development events during metamorphic climax. The pattern of timing of chondrification of the anterior nasal cartilages more closely coincides with that of the neobatrachian species than that recorded for the pelobatid frog Spea. In addition, this study supports a sister taxon relationship between Discoglossus and Alytes.

Pelvic and thigh musculature in frogs (Anura) and origin of anuran jumping locomotion.

Comparative analysis of the anuran pelvic and thigh musculoskeletal system revealed that the thigh extensors, responsible for the initial phase of jump, the propulsive stroke in swimming and, if used asynchronously, also for walking, are least affected by the transformations observed between anurans and their temnospondyl ancestors (as reflected in contemporary caudates). The iliac shaft and urostyle, two of the most important anuran apomorphies, represent skeletal support for muscles that are mostly protractors of the femur or are important in attaining a crouching position, a necessary prerequisite for rapid escape. All of these muscles originate or insert on the iliac shaft. As the orientation of the pubis, ischium and ilium is the same in anurans, caudates and by inference also in their temnospondyl ancestors, it is probable that the pelvis was shifted from the sacral vertebra posteriorly along the reduced and stiffened tail (urostyle) by the elongation of the illiac shaft. Thus, the original vertical orientation of the ilium was maintained (which is also demonstrated by stable origins of the glutaeus maximus, iliofemoralis and iliofibularis on the tuber superius) and the shaft itself is a new structure. A review of functional analysis of anuran locomotion suggests some clear differences from that in caudates, suggesting that terrestrial jumping may have been a primary locomotor activity, from which other types of anuran locomotion are derived.

The evolution of amphibian metamorphosis: insights based on the transformation of the aortic arches of Pelobates fuscus (Anura).

In order to gain insights into how the aortic arches changed during the transition of vertebrates to land, transformations of the aortic arches during the metamorphosis of Pelobates fuscus were investigated and compared with data from the early development of a recent ganoid fish Amia calva and a primitive caudate amphibian Salamandrella keyserlingi. Although in larval Pelobates, as in other non-pipid anurans, the gill arches serve partly as a filter-feeding device, their aortic arches maintain the original piscine-like arrangement, except for the mandibular and hyoid aortic arches which were lost. As important pre-adaptations for breathing of atmospheric oxygen occur in larval Pelobates (which have well-developed, though non-respiratory lungs and pulmonary artery), transformation of aortic arches during metamorphosis is fast. The transformation involves disappearance of the ductus Botalli, which results in a complete shunting of blood into the lungs and skin, disappearance of the ductus caroticus, which results in shunting of blood into the head through the arteria carotis interna, and disappearance of arch V, which results in shunting blood to the body through arch IV (systemic arch). It is supposed that the branching pattern of the aortic arches of permanently water-dwelling piscine ancestors, of intermediate forms which occasionally left the water and of primitive tetrapods capable of spending longer periods of time on land had been the same as in the prematamorphic anuran larvae or in some metamorphosed caudates in which the ductus caroticus and ductus Botalli were not interrupted, and arch V was still complete.

Transformation of the pectoral girdle in the evolutionary origin of frogs: insights from the primitive anuran Discoglossus.

Using cleared-and-stained whole mounts and computer-aided three-dimensional reconstructions made from serial histological sections, we studied the development of the pectoral girdle in Discoglossus pictus, an extant member of an ancient frog lineage, represented for example by Eodiscoglossus from the Middle Jurassic to Early Cretaceous periods in Europe. Basic developmental features were compared with those of extinct Temnospondyli, considered to be the most probable anuran ancestors, and with Triadobatrachus, an early Triassic proanuran. In the endochondral girdle, the separate scapula and coracoid of Discoglossus and other anurans (completed by suprascapular and procoracoid cartilages) evolved from the compact scapulocoracoid of temnospondyls by paedomorphosis. In parallel, the dermal ossifications of the girdle were reduced to a small clavicle and cleithrum. The overall reduction in ossification of the anuran pectoral girdle supports the hypothesis of a paedomorphic origin for Anura. The almost simultaneous appearance of dermal and endochondral ossifications may be explained by the accumulation of developmental events during a short, distinct metamorphosis (which did not occur in neotenic temnospondyls living permanently in water). The sternal elements seem to be neomorphs for the most part, which help to cushion the shock of landing in jumping anurans but which also evolved as functional substitutes (insertion area for the pectoralis muscles) of the temnospondyl interclavicle.

Development of the pelvis and posterior part of the vertebral column in the Anura.

The anuran pelvic girdle is unique among all amphibians in that its acetabular portion is located far posterior to the sacrum, lateral to the postsacral (= caudal) vertebral column, which is reduced to a single rod-like element called the urostyle. This situation in the adult is strikingly different not only from that in ancestral temnospondyls but also in other modern amphibians. Because there is no fossil that would document this evolutionary anatomical modification except for Triadobatrachus, the only data may be inferred from development in modern anurans. We chose seven anuran species (belonging to the genera Discoglossus, Bombina, Pelobates, Bufo, Rana and Xenopus), representing the principal locomotory types (saltation, swimming, crawling and burrowing). Development of the pelvic girdle was studied on cleared and stained whole mounts and partly on serial histological sections. The basic developmental pattern was similar in all species: the pelvis on both sides develops from two centres (puboischiadic and iliac, respectively). The ilium then extends vertically towards the sacral vertebra and later rotates posteriorly so that ultimately the acetabulum is lateral to the tail (= urostyle). Only minor deviations from this pattern were found, mainly associated with differences in water and terrestrial dwelling.

Developmental origin of the frontoparietal bone in Bombina variegata (Anura: Discoglossidae).

Development of the frontoparietal bone in the European yellow-bellied toad, Bombina variegata, was followed on the basis of histological analysis of transverse serial sections through the larval skulls to recognize early stages of ossification represented by osteoid (uncalcified bone matrix) and on cleared and stained specimens to investigate more advanced stages. Ossification of the frontal begins as three tiny areas of osteoid (F(1), F(2), F(3)) adjoining the dorsal surface of the orbital cartilage, which are separated by areas without osteoid. F(3) is the largest (most advanced). Prior to calcification, F(3) extends to fuse with F(2) and then with F(1), but it does not expand over the prootic fissure posteriorly. As calcification begins the strip of bone is joined posteromedially by F(4). Only then does a single ossification center corresponding to the parietal arise on the anterodorsal surface of the otic capsule. This ossification sequence corresponds to those observed in the Actinopterygii and in caudate amphibians.

Skull of the neotenic salamandrid amphibian Triturus alpestris and abbreviated development in the Tertiary Salamandridae.

The skull of neotenic individuals of the Alpine Newt Triturus alpestris from the locality Drakolimni (Greece) is described on the basis of models made from magnified serial frontal sections. In order to recognize features associated with neoteny and paedomorphosis, the results were compared with normal development of contemporary Triturus and of other Caudata. The neotenic larvae from Greece correspond to advanced stages of metamorphosis in normal development of Triturus alpestris. Comparison with salamandrids from the Tertiary of Europe in which the hyobranchial skeleton was preserved, namely Brachycormus noachicus, Chelotriton paradoxus, and Palaeopleurodeles hauffi, revealed that both latter taxa were completely metamorphosed adults anatomically similar to their contemporary relatives Tylototriton, Echinotriton, and Pleurodeles, whereas Brachycormus, though apparently related to Chelotriton, was a neotenic amphibian. This is suggested by its incompletely metamorphosed but ossified hyobranchial skeleton. © 1996 Wiley-Liss, Inc.

Development of the ethmoidal structures of the endocranium in the anuran Pipa pipa.

Investigation of eight developmental stages by means of serial sections and subsequent graphic or wax model reconstructions, as well as by means of cleared-and-stained and dissected material, revealed that the ethmoidal endocranium in Pipa pipa consists in early states of a single horizontal ethmoid plate lacking labial cartilages. Later in the course of development, structures comparable with those in other anurans appear, though modified and of reduced size. These adult structures arise from the new cartilaginous tissue located above the former larval ethmoid plate, whereas the latter entirely disappear. This phenomenon can be observed also in P. carvalhoi and in Xenopus laevis; hence, it supposedly occurs in all pipids. On the other hand, in anuran larvae, which develop cornua trabecularum in the ethmoidal region, these persist in adults as part of the nasal septum. Positional and developmental differences suggest that, although the ethmoid plate and the cornua trabecularum arise from the same region of the cranial neural crest, they are not fully corresponding structures. Comparison with adults of other pipid genera confirmed the conclusion of some earlier investigators that P. pipa is the most specialized among pipids.